Compounds, Compositions, and Methods for use in Treating Autophagy-Associated Disorders

ABSTRACT

The invention provides compounds and methods of treating autophagy mediated diseases and disorders and related pharmaceutical compositions, diagnostics, screening techniques and kits.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional application of U.S. Non-Provisionalapplication Ser. No. 16/717,728, titled “Compounds, Composition, andMethods for use in Treating Autophagy-Associated Disorders,” filed Dec.17, 2019, which claims priority to U.S. Provisional Application No.62/780,615, titled “Compositions and Methods for use in TreatingAutophagy-Associated Disorders,” filed Dec. 17, 2018, the contents ofwhich are incorporated by reference herein in their entirety.

FIELD OF THE INVENTION

The invention provides compounds, compositions, and methods for treatingautophagy-associated disorders and related pharmaceutical compositions,diagnostics, screening techniques and kits.

BACKGROUND

Autophagy is a homeostatic process that is highly conserved ineukaryotic cells, where it acts as a cytoplasmic biomass quantity andquality control system (Mizushima N., et al., Nature (2008) 451,1069-1075; Yang Z., and Klionsky, D. J., Nat. Cell Biol. (2010) 12,814-822). Autophagy functions encompass programmed cell survival andcell death, normally skewed toward cell survival (Kroemer G. and Levine,B., Nat Rev Mol Cell Biol (2008) 9, 1004-1010) through provision ofenergy and nutrients and ridding the cytoplasm of toxic macromolecularaggregates, faulty organelles (Mizushima, et al., 2008; Yang Z., andKlionsky, D. J., 2010) and invading microorganisms (Deretic, V., andLevine, B., Cell Host Microbe (2009) 5, 527-549; Levine, B., et al.,Nature (2011) 469, 323-335).

The cell-autonomous antimicrobial defense functions of autophagy,demonstrated initially in the case of Streptococci (Nakagawa, I., etal., Science (2004) 306, 1037-1040) and Mycobacterium tuberculosis(Gutierrez, M. G., et al., Cell (2004) 119, 753-766; Harris, J., et al.,Immunity (2007) 27, 505-517; Ponpuak, M., et al., Immunity (2010) 32,329-341), have been extended to a wide variety of microbes with a caveatthat most highly adapted pathogens have evolved specific protectivemechanisms against autophagic elimination of microbes (Deretic andLevine, 2009; Gannage M. et al., Cell Host Microbe (2009) 6, 367-380;Kyei G. B. et al., J Cell Biol (2009) 186, 255-268; Lee J. S., et al.,Nat Cell Biol (2009) 11, 1355-1362; Orvedahl A. et al., Cell Host andMicrobe (2007) 1, 23-35; Yoshikawa, Y., et al. Nat Cell Biol (2009) 11,1233-1240.). Other studies have uncovered orderly intersections betweenautophagy and innate immunity (Chaturvedi, A., et al., Dorward, D., andPierce, S. K., Immunity (2008) 28, 799-809; Cooney, R., et al., Nat Med(2010) 16, 90-97; Delgado, M. A., et al., Embo J (2008) 27, 1110-1121;Huang, J., Proc Natl Acad Sci USA (2009) 106(15):6226-31; Sanjuan, M.A., et al., Nature (2007) 450, 1253-1257; Shi, C. S., and Kehrl, J. H.,Sci Signal (2010) 3, ra42; Singh, S. B., Davis, A. S., Taylor, G. A.,and Deretic, V., Science (2006) 313, 1438-1441; Tang, D., et al., J CellBiol (2010) 190, 881-892; Travassos, L. H., et al. Nat Immunol (2010)11, 55-62; Xu, Y., Jagannath, C., et al., Immunity (2007) 27, 135-144;Yano, T., Mita, S., Ohmori, H., Oshima, Y., Fujimoto, Y., Ueda, R.,Takada, H., Goldman, W. E., Fukase, K., Silverman, N., et al., NatImmunol (2008) 9, 908-916.), adaptive immunity (Blanchet, F. P., et al.,Immunity (2010) 32, 654-669; Lee, H. K., et al., Immunity (2010) 32,227-239; Munz, C. (2009). Enhancing immunity through autophagy. Annu RevImmunol 27, 423-449; Nedjic, J., et al., Nature (2009) 455, 396-400;Paludan, C., et al., Science (2005) 307, 593-596), T-cell development,differentiation and homeostasis (Jia, W., and He, Y. W., J Immunol(2011) 186, 5313-5322; Johansen, T., and Lamark, T. (2011).

Autophagy (2011) 7; Nedjic, J., et al., Nature (2008) 455, 396-400), andinflammatory responses (Cadwell, K., et al., Cell (2010) 141, 1135-1145;Jounai, N., et al., Proc Natl Acad Sci USA (2007) 104, 14050-14055;Levine B, et al., Nature (2011) 469: 323-335; Saitoh, T., and Akira, S.,J Cell Biol (2010) 189, 925-935). It was also found that autophagysuppresses endogenous, cell-autonomous promoters of inflammation(Mathew, R., et al., Cell (2009) 137, 1062-1075; Orvedahl, A., et al.,Cell Host Microbe (2010) 7, 115-127).

Specific autophagic factors, such as Atg5-Atg12, have been shown toinhibit RIG-I signaling (Jounai, N., et al., Proc Natl Acad Sci USA(2007) 104, 14050-14055) whereas Atg9 has been reported to negativelyregulate trafficking, assembly and activation of TBK-1 (TANK-bindingkinase 1), which, among its key functions, controls type I interferonresponse elicited by intracellular double stranded DNA (Saitoh, T., etal., Proc Natl Acad Sci USA (2009) 106, 20842-20846). In the context ofanti-inflammatory function, recent studies indicate that autophagy playsan inhibitory role in inflammasome and IL-1β activation by mechanismsthat involve mitochondrial homeostasis (Nakahira K, et al., Nat Immunol(2010) 12: 222-230; Zhou R, et al., Nature (2011) 469: 221-225) orpotentially direct effects (Harris J, et al., J Biol Chem (2011) 286:9587-9597). Finally, a number of genetic links have been found in humanpopulations between autophagy and idiopathic inflammation (Consortium,Nature (2007) 447, 661-678; Craddock, N., et al. Nature (2010) 464,713-720.) or infectious diseases such as tuberculosis (Che, N., et al.,Clin Chim Acta (2010) 411, 1645-1649; Intemann, C. D., et al., PLoSPathog (2009) 5, e1000577; Singh, S. B., et al., Science (2006) 313,1438-1441; Singh, S. B., et al., Nat Cell Biol (2010) 12, 1154-1165),with significant inflammatory components and tissue damage.

Given the interconnectedness of autophagy and immunity, it is likelythat the immune manifestations of autophagy are affected not only by theinduction of autophagy but also by the completion of the autophagicpathway. The formation of the autophagic organelles of the sensu strictoautophagy pathway (also referred to as macroautophagy) depends onmultiple sources of membrane or regulatory factors (Tooze, S. A., andYoshimori, T., Nat Cell Biol (2010) 12, 831-835). The key stages ofautophagy, however, are not restricted to the formation ofautophagosomal membranes and include the sequestration of the earmarkedcargo by the autophagic adaptors (Bjorkoy, G., et al., J Cell Biol(2005) 171, 603-614; Kirkin, V., et al. Mol Cell (2009) 33, 505-516;Thurston, T. L., et al., Nat Immunol (2009) 10, 1215-1221; Wild, P., etal., Science (2011) 333, 228-233), and the less understood process ofthe maturation of autophagic organelles into autolysosomes where thecaptured material is degraded (Liang, C., et al., Nat Cell Biol (2008)10, 776-787; Matsunaga, K., et al. Nat Cell Biol (2009) 11, 385-396).

Autophagy is directly implicated in cancer, type II diabetes,neurodegenerative syndromes such as Alzheimer's, Huntington's andParkinson's diseases, chronic inflammatory diseases (e.g. Crohn'sdisease), type II diabetes, infections (such as tuberculosis and HIV (Iand II)/AIDS, hepatitis B, hepatitis C), and a variety of disordersassociated with aging. A better understanding of how autophagicmechanisms are implicated in the aforementioned diseases could provecritical to preventing or treating these maladies.

SUMMARY

The elucidation of certain autophagic processes involved in the onsetand progression of a variety of infectious, inflammatory, developmental,chronic pain, and depression-related disorders has led to the discoveryof compounds and methods to treat and diagnose such ailments. Further,the compounds are effective as modulators of autophpagy in the treatmentof infectious, inflammatory, developmental, chronic pain, anddepression-related disorders, as well as being used in analyses ofautophagic processes, including the disease state in a patient fordiagnosis and/or monitoring therapy of the disease state. The presentinvention is directed to compounds which exhibit biological activity asmodulators (inhibitors or agonists) of autophagy and consequently can beused in the treatment of diseases which occur or are mediated throughautophagy.

In one embodiment, the present invention provides novel compounds. Thecompounds can be used to treat a subject, for example, to modulateautophagy. In this aspect of the invention, a compound identified hereinas an autophagy modulator (inhibitor or agonist, also referred togenerically as an “autostatin”) is presented to the biological system,including administration to a patient or subject in need, in order tomodulate (often enhance or up-regulate but in certain instances, toinhibit) autophagy and effect a favorable result in the biologicalsystem. The resulting modulation may be monitored or applied in thebiological system to effect a favorable result, including theinhibition, treatment and/or prevention of cancer, including metastasisof cancer, or the inhibition, treatment (including the amelioration ofsymptoms), and/or prevention of one or more disease states or conditionsin which the modulation, especially including upregulation or inhibitionof autophagy provides a favorable result in numerous disease statesand/or conditions including neurodegeneration (including, for example,Alzheimer's disease, Parkinson's disease; other ataxias), chronicinflammatory diseases (including, for example, inflammatory boweldisease, including Crohn's disease, rheumatoid arthritis, lupus,multiple sclerosis, chronic obstructive pulmonary disease/COPD,pulmonary fibrosis, cystic fibrosis, Sjogren's disease), diabetes andmetabolic syndrome, muscle degeneration and atrophy, frailty in aging,stroke and spinal cord injury, arteriosclerosis, infectious diseases(HIV, HBV, HCV, including secondary associated disease states orconditions, including AIDS) and tuberculosis, among others, and chronicpain related disorders (including, for example, chronic joint pain,chronic back pain, chronic nerve pain, chronic headaches, trigeminalpain, myofascial pain syndrome/MPS, fibromyalgia), and depressiveconditions (including, for example, post-traumatic stress disorder/PTSD,chronic pain syndrome/CPS, chronic regional pain syndrome/CRPS) andrelated disorders, and developmental diseases (both overly mature andimmature development, including erythrocyte differentiation,embryogenesis/fertility and ageing/progeria). The common principle ofthis embodiment of the invention is that compounds, includingautostatins which are autophagy modulators, depending upon the diseasestate, condition or symptom to be treated, may cure, prevent (includingreducing the likelihood of), improve prognosis, ameliorate symptomsand/or improve the quality of the patient's or subject's life. Inaddition, in the therapeutic aspects of the invention, theadministration of the compound of the invention may prolong the life ofthe patient, as well as improve the quality of life in the aging patientor subject.

The compound of the invention can be used to treat an autophagy-mediateddisease alone or in combination with at least one autostatin orbioactive agent. An autostatin such as, for example, flubendazole,hexachlorophene, propidium iodide, bepridil, clomiphene citrate (Z,E),GBR 12909, propafenone, metixene, dipivefrin, fluvoxamine, dicyclomine,dimethisoquin, ticlopidine, memantine, bromhexine, norcyclobenzaprine,diperodon, nortriptyline, tetrachlorisophthalonitrile and phenylmercuricacetate, benzethonium, niclosamide, monensin, bromperidol, levobunolol,dehydroisoandosterone 3-acetate, sertraline, tamoxifen, reserpine,hexachlorophene, dipyridamole, harmaline, prazosin, lidoflazine,thiethylperazine, dextromethorphan, desipramine, mebendazole, canrenone,chlorprothixene, maprotiline, homochlorcyclizine, loperamide,nicardipine, dexfenfluramine, nilvadipine, dosulepin, biperiden,denatonium, etomidate, toremifene, tomoxetine, clorgyline, zotepine,beta-escin, tridihexethyl, ceftazidime, methoxy-6-harmalan,melengestrol, albendazole, rimantadine, chlorpromazine, pergolide,cloperastine, prednicarbate, haloperidol, clotrimazole, nitrofural,iopanoic acid, naftopidil, methimazole, trimeprazine, ethoxyquin,clocortolone, doxycycline, pirlindole mesylate, doxazosin, deptropine,nocodazole, scopolamine, oxybenzone, halcinonide, oxybutynin,miconazole, clomipramine, cyproheptadine, doxepin, dyclonine,salbutamol, flavoxate, amoxapine, fenofibrate, pimethixene,pharmaceutically acceptable salts thereof and mixtures thereof, can beadministered to a patient or subject in need to treat anautophagy-mediated disease state and/or condition. The autostatin can beeither an agonist or inducer of autophagy, or anantagonist or inhibitorof autophagy. The compounds of the invention can be used as modulatorsof autophagy in the various autophagy-mediated disease states andconditions described herein, with the agonists being preferred in mostdisease states other than cancer and in the case of the treatment ofcancer, the inhibitors described above are preferred, alone or incombination with an autophagy agonist as described above and/or anadditional anticancer agent as otherwise described herein.

The additional bioactive agent can be, for example, an anticancer agent,antibiotic, anti-tuberculosis agent, antiviral agent such as an anti-HIVagent, anti-HBV agent or anti-HCV agent. Additionally, at least oneanticancer agent can be administered in combination with the compound ofthe invention. In the present invention, an autostatin may be combinedwith an additional autophagy modulator.

The present invention also relates to compounds which can be used forthe treatment of an autophagy mediated disease state or condition. Thus,the present invention is also directed to pharmaceutical compositionswhich comprise an effective amount of at least one compound describedherein, either alone or in combination with a pharmaceuticallyacceptable carrier, additive or excipient, an autostatin, at least oneadditional bioactive agent. Such disease states or conditions, include,for example, cancer, including metastasis of cancer, lysosomal storagediseases, neurodegeneration (including, for example, Alzheimer'sdisease, Parkinson's disease; other ataxias), immune response, chronicinflammatory diseases, including inflammatory bowel disease, includingCrohn's disease, rheumatoid arthritis, lupus, multiple sclerosis,chronic obstructive pulmonary disease/COPD, pulmonary fibrosis, cysticfibrosis, Sjogren's disease; diabetes (I and II) and metabolic syndrome,liver disease, renal disease (including glomerular disease),cardiovascular disease (especially including ischemia, stroke, pressureoverload and complications during reperfusion), muscle degeneration andatrophy, frailty in aging, stroke and spinal cord injury,arteriosclerosis, infectious diseases (microbial infections, includingbacterial, fungal, cellular and viral, including secondary diseasestates or conditions associated with infectious diseases), includingAIDS and tuberculosis, among others, developmental disease (both overlymature and immature development, including erythrocyte differentiation,embryogenesis/fertility and ageing/progeria), and chronic pain relateddisorders (including, for example, chronic joint pain, chronic backpain, chronic nerve pain, chronic headaches, trigeminal pain, myofascialpain syndrome/MPS, fibromyalgia), and depressive conditions (including,for example, post-traumatic stress disorder/PTSD, chronic painsyndrome/CPS, chronic regional pain syndrome/CRPS) and relateddisorders. The common principle of this embodiment of the invention isthat compounds of the invention may be used for the treatment of thedisease state and/or condition which is mediated through autophagy.

DETAILED DESCRIPTION

It is noted that, as used in this specification and the appended claims,the singular forms “a,” “an,” and “the,” include plural referents unlessexpressly and unequivocally limited to one referent. Thus, for example,reference to “a compound” includes two or more different compound. Asused herein, the term “include” and its grammatical variants areintended to be non-limiting, such that recitation of items in a list isnot to the exclusion of other like items that can be substituted orother items that can be added to the listed items.

The term “compound” or “agent,” as used herein, refers to any specificchemical compound disclosed herein and includes tautomers, regioisomers,geometric isomers as applicable, and also where applicable, opticalisomers (e.g. enantiomers) thereof, as well as pharmaceuticallyacceptable salts thereof. Within its use in context, the term compoundgenerally refers to a single compound, but also may include othercompounds such as stereoisomers, regioisomers and/or optical isomers(including racemic mixtures) as well as specific enantiomers orenantiomerically enriched mixtures of disclosed compounds as well asdiastereomers and epimers, where applicable in context. The term alsorefers, in context to prodrug forms of compounds which have beenmodified to facilitate the administration and delivery of compounds to asite of activity.

The term “patient” or “subject” is used throughout the specificationwithin context to describe an animal, generally a mammal and preferablya human, to whom treatment, including prophylactic treatment(prophylaxis), with the methods and compositions according to thepresent invention is provided. For treatment of those conditions ordisease states which are specific for a specific animal such as a humanpatient, the term patient refers to that specific animal, often a human.

The terms “effective” or “pharmaceutically effective” are used herein,unless otherwise indicated, to describe an amount of a compound orcomposition which, in context, is used to produce or affect an intendedresult, usually the modulation of autophagy within the context of aparticular treatment or alternatively, the effect of a bioactive agentwhich is coadministered with the autophagy modulator (autotoxin) in thetreatment of disease.

The terms “treat,” “treating,” and “treatment,” as used herein, refer toany action providing a benefit to a patient at risk for or afflicted byan autophagy mediated disease state or condition. The benefit may be incuring the disease state or condition, inhibition of its progression, orameliorating, lessening or suppressing one or more symptoms of anautophagy mediated disease state or condition. Treatment encompassesboth prophylactic and therapeutic treatment.

As used herein, the term “autophagy mediated disease state or condition”refers to a disease state or condition that results from disruption inautophagy or cellular self-digestion. Autophagy is a cellular pathwayinvolved in protein and organelle degradation, and has a large number ofconnections to human disease. Autophagic dysfunction is associated withcancer, neurodegeneration, microbial infection and ageing, amongnumerous other disease states and/or conditions. Although autophagyplays a principal role as a protective process for the cell, it alsoplays a role in cell death. Disease states and/or conditions which aremediated through autophagy (which refers to the fact that the diseasestate or condition may manifest itself as a function of the increase ordecrease in autophagy in the patient or subject to be treated andtreatment requires administration of an inhibitor or agonist ofautophagy in the patient or subject) include, for example, cancer,including metastasis of cancer, lysosomal storage diseases,neurodegeneration (including, for example, Alzheimer's disease,Parkinson's disease; other ataxias), immune response (T cell maturation,B cell and T cell homeostasis, counters damaging inflammation) andchronic inflammatory diseases (may promote excessive cytokines whenautophagy is defective), including, for example, inflammatory boweldisease, including Crohn's disease, rheumatoid arthritis, lupus,multiple sclerosis, chronic obstructive pulmonary disease/COPD,pulmonary fibrosis, cystic fibrosis, Sjogren's disease, hyperglycemicdisorders, diabetes (I and II), affecting lipid metabolism isletfunction and/or structure, excessive autophagy may lead to pancreaticβ-cell death and related hyperglycemic disorders, including severeinsulin resistance, hyperinsulinemia, insulin-resistant diabetes (e.g.Mendenhall's Syndrome, Werner Syndrome, leprechaunism, and lipoatrophicdiabetes) and dyslipidemia (e.g. hyperlipidemia as expressed by obesesubjects, elevated low-density lipoprotein (LDL), depressed high-densitylipoprotein (HDL), and elevated triglycerides) and metabolic syndrome,liver disease (excessive autophagic removal of cellularentities-endoplasmic reticulum), renal disease (apoptosis in plaques,glomerular disease), cardiovascular disease (especially includingischemia, stroke, pressure overload and complications duringreperfusion), muscle degeneration and atrophy, frailty in aging, strokeand spinal cord injury, arteriosclerosis, infectious diseases (microbialinfections, removes microbes, provides a protective inflammatoryresponse to microbial products, limits adaptation of autophagy of hostby microbe for enhancement of microbial growth, regulation of innateimmunity) including bacterial, fungal, cellular and viral (includingsecondary disease states or conditions associated with infectiousdiseases), including AIDS and tuberculosis, among others, and chronicpain related disorders (including, for example, chronic joint pain,chronic back pain, chronic nerve pain, chronic headaches, trigeminalpain, myofascial pain syndrome/MPS, fibromyalgia), and depressiveconditions (including, for example, post-traumatic stress disorder/PTSD,chronic pain syndrome/CPS, chronic regional pain syndrome/CRPS) andrelated disorders. development (including erythrocyte differentiation),embryogenesis/fertility/infertility (embryo implantation and neonatesurvival after termination of transplacental supply of nutrients,removal of dead cells during programmed cell death) and ageing(increased autophagy leads to the removal of damaged organelles oraggregated macromolecules to increase health and prolong life, butincreased levels of autophagy in children/young adults may lead tomuscle and organ wasting resulting in ageing/progeria).

The term “lysosomal storage disorder” refers to a disease state orcondition that results from a defect in lysosomal storage. These diseasestates or conditions generally occur when the lysosome malfunctions.Lysosomal storage disorders are caused by lysosomal dysfunction, usuallyas a consequence of deficiency of a single enzyme required for themetabolism of lipids, glycoproteins or mucopolysaccharides. Theincidence of lysosomal storage disorder (collectively) occurs at anincidence of about 1:5,000-1:10,000. The lysosome is commonly referredto as the cell's recycling center because it processes unwanted materialinto substances that the cell can utilize. Lysosomes break down thisunwanted matter via high specialized enzymes. Lysosomal disordersgenerally are triggered when a particular enzyme exists in too small anamount or is missing altogether. When this happens, substancesaccumulate in the cell. In other words, when the lysosome doesn'tfunction normally, excess products destined for breakdown and recyclingare stored in the cell. Lysosomal storage disorders are geneticdiseases, but these may be treated using autophagy modulators asdescribed herein.

Examples of lysosomal storage disease include, for example, activatordeficiency/GM2 gangliosidosis, alpha-mannosidosis,aspartylglucoaminuria, cholesteryl ester storage disease, chronichexosaminidase A deficiency, cystinosis, Danon disease, Fabry disease,Farber disease, fucosidosis, galactosialidosis, Gaucher Disease (TypesI, II and III), GM1 Ganliosidosis, including infantile, lateinfantile/juvenile and adult/chronic), Hunter syndrome (MPS II), I-Celldisease/Mucolipidosis II, Infantile Free Sialic Acid Storage Disease(ISSD), Juvenile Hexosaminidase A Deficiency, Krabbe disease, Lysosomalacid lipase deficiency, Metachromatic Leukodystrophy, Hurler syndrome,Scheie syndrome, Hurler-Scheie syndrome, Sanfilippo syndrome, MorquioType A and B, Maroteaux-Lamy, Sly syndrome, mucolipidosis, multiplesulfate deficiency, Niemann-Pick disease, Neuronal ceroidlipofuscinoses, CLN6 disease, Jansky-Bielschowsky disease, Pompedisease, pycnodysostosis, Sandhoff disease, Schindler disease, Tay-Sachsand Wolman disease, among others.

The term “modulator of autophagy,” “regulator of autophagy” or“autostatin” is used to refer to a compound which functions as anagonist (inducer or up-regulator) or antagonist (inhibitor ordown-regulator) of autophagy. Depending upon the disease state orcondition, autophagy may be upregulated (and require inhibition ofautophagy for therapeutic intervention) or down-regulated (and requireupregulation of autophagy for therapeutic intervention). In mostinstances, in the case of cancer treatment with a modulator of autophagyas otherwise described herein, the autophagy modulator is often anantagonist of autophagy. In the case of cancer, the antagonist(inhibitor) of autophagy may be used alone or combined with an agonistof autophagy.

The compounds of the invention can be used in the treatment of anautophagy mediated disease state or condition as otherwise describedherein. The following compounds have been identified and the structuresare set forth below.

The following compounds share the following backbone:

-   -   a)        2,6-di(3,4-methylenedioxyphenyl)-4-(2-bromo-4,5-methylenedioxyphenyl)pyridine    -   b) 2,6-di(3,4-methylenedioxyphenyl)-4-(2-methoxyphenyl)pyridine    -   c) 2,6-di(2-methoxyphenyl)-4-(4-methoxyphenyl) pyridine    -   d) 2,6-di(2-methoxyphenyl)-4-(4-hydroxyphenyl)pyridine    -   e)        2,6-di(2-methoxyphenyl)-4-(2-bromo-4,5-methylenedioxyphenyl)pyridine    -   f) 2,6-di(2-methoxyphenyl)-4-(3-methoxyphenyl)pyridine    -   g) 2,6-di(2-methoxyphenyl)-4-(4-acetomidophenyl)pyridine    -   h) 2,6-di(2-methoxyphenyl)-4-(4-cyanophenyl) pyridine    -   i) 2,6-di(2-methoxyphenyl)-4-(3-cyanophenyl)pyridine    -   j) 2,6-di(2-methoxyphenyl)-4-(4-carboxyphenyl)pyridine    -   k) 2,6-di(2-methoxyphenyl)-4-(3,4-dimethoxyphenyl)pyridine    -   l) 2,6-di(2-methoxyphenyl)-4-(dimethylaminophenyl)pyridine    -   m) 2-(2-methoxyphenyl)-6-(3-methoxyphenyl)-4-(4-methoxyphenyl)        pyridine    -   n) 2,6-di(4-methoxyphenyl)-4-(4-methoxyphenyl)pyridine    -   o) 2,6-di(3-methoxyphenyl)-4-(4-methoxyphenyl)pyridine    -   p) 2,6-di(2-hydroxyphenyl)-4-(4-hydroxyphenyl)pyridine    -   q) 2,6-di(2,4-dimethoxyphenyl)-4-(4-hydroxyphenyl)pyridine    -   r) 2,6-di(2,4-dimethoxyphenyl)-4-(2,5-dihydroxyphenyl)pyridine    -   s)        2-(3,4-methylenedioxy)-6-(3-methoxyphenyl)-4-(4-methoxyphenyl)        pyridine    -   t) 2-(3-methoxyphenyl)-6-(4-methoxyphenyl)-4-(4-methoxyphenyl)        pyridine    -   u)        2-(3,4-methylenedioxy)-6-(2-methoxyphenyl)-4-(4-methoxyphenyl)        pyridine

The following compounds share the following backbone:

-   -   a)        N-(3,4-dimethoxypheny)-1,8-Dioxa-3-aza-2,4-dihydro-2H-5-phenyl-anthracen-7-one    -   b)        N-(3,4-dimethoxybenzyl)-1,8-Dioxa-3-aza-2,4-dihydro-2H-5-methyl-anthracen-7-one    -   c) N-benzyl-1,8-Dioxa-3-aza-3,4dihydro-2H-anthracen-7-one series    -   d)        N-(3,4-dimethoxypheny)-1,8-Dioxa-3-aza-2,4-dihydro-2H-5-methyl-anthracen-7-one

The following compounds share the following backbone:

-   -   a)        7-hydoxy-3-(4-(3,4-dimethoxybenzyl))-3,4-dihydro-2H-1,3-benzoxazine    -   b)        5,6-dimethoxy-3-(4-methoxybenzyl)-3,4-dihydro-2H-1,3-benzoxazine    -   c)        7-Iodo-3-(4-(3,4-dimethoxybenzyl))-3,4-dihydro-2H-1,3-benzoxazine    -   d)        6-Methoxy-3-(4-(3,4-dimethoxybenzyl))-3,4-dihydro-2H-1,3-benzoxazine    -   e)        6-Methoxy-5,7,8-trimethyl-3-(4-(3,4-dimethoxybenzyl))-3,4-dihydro-2H-1,3-benzoxazine    -   f)        5,6-methylenedioxy-3-(4-methoxybenzyl)-3,4-dihydro-2H-1,3-benzoxazine    -   g)        6-Trifluoromethoxy-3-(4-(3,4-dimethoxybenzyl))-3,4-dihydro-2H-1,3-benzoxazine    -   h) 7-Hydroxy-3-(4-hydroxyphenyl)-3,4-dihydro-2H-1,3-benzoxazine    -   i) 6-Methoxy-3-(4-methoxyphenyl)-3,4-dihydro-2H-1,3-benzoxazine    -   j)        5-Hydroxy-6,8-dichloro-3-(4-trifluoromethoxyphenyl)-3,4-dihydro-2H-1,3-benzoxazine    -   k)        5,7-dihydroxy-3-(4-hydroxyphenyl)-3,4-dihydro-2H-1,3-benzoxazine    -   l)        6-hydroxy-3-(2-chloro-4-ihydroxyphenyl)-3,4-dihydro-2H-1,3-benzoxazine    -   m)        6-Iodo-3-(2,chloro-4-iodophenyl)-3,4-dihydro-2H-1,3-benzoxazine    -   n)        5,6-dimethoxy-3-(4-methoxyphenyl)-3,4-dihydro-2H-1,3-benzoxazine    -   o)        5,6-dimethoxy-3-(4-methoxyphenyl)-3,4-dihydro-2H-1,3-benthiazine

The following compounds share the following backone:

-   -   a) 5-Chloro-2-hydroxy-N-(3-hydroxy 4-methoxy)benzylbenzamide    -   b) 5-Chloro-2-hydroxy-N-(2-chloro-4-trifluoromethyl)        phenylbenzamide    -   c) 5-Chloro-2-hydroxy-N-(3,4,5-trimethoxy)phenylbenzamide

The following compounds share the following backone:

-   -   a) 3-(4-methoxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione    -   b) 3-(4-hydroxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione    -   c) 3-(4-methoxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione    -   d)        3-(4-allyloxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione    -   e) 3-(4-hydroxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione    -   f)        3-(3,4,5-trimethoxymethoxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione    -   g)        3-(4-allyloxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione    -   h)        3-(4-dimethylaminophenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione

These compounds, their analogs and pharmaceutically acceptable saltsthereof can be used as modulators of autophagy in variousautophagy-mediated disease states and conditions described herein, withthe agonists being preferred in most disease states other than cancer(although inhibitors may also be used alone, or preferably incombination with the agonists) In the case of the treatment of cancer,the inhibitors described above are preferred, alone or in combinationwith an autophagy agonist as described above and/or an additionalanticancer agent as otherwise described herein.

Other compounds which may be used in combination with the compounds ofthe invention include, for example, other “additional autophagymodulators” or “additional autostatins” which are known in the art.These can be combined with one or more of the compounds to provide novelpharmaceutical compositions and/or methods of treating autophagymediated disease states and conditions which are otherwise describedherein. These additional autophagy modulators include, for example,benzethonium, niclosamide, monensin, bromperidol, levobunolol,dehydroisoandosterone 3-acetate, sertraline, tamoxifen, reserpine,hexachlorophene, dipyridamole, harmaline, prazosin, lidoflazine,thiethylperazine, dextromethorphan, desipramine, mebendazole, canrenone,chlorprothixene, maprotiline, homochlorcyclizine, loperamide,nicardipine, dexfenfluramine, nilvadipine, dosulepin, biperiden,denatonium, etomidate, toremifene, tomoxetine, clorgyline, zotepine,beta-escin, tridihexethyl, ceftazidime, methoxy-6-harmalan,melengestrol, albendazole, rimantadine, chlorpromazine, pergolide,cloperastine, prednicarbate, haloperidol, clotrimazole, nitrofural,iopanoic acid, naftopidil, Methimazole, Trimeprazine, Ethoxyquin,Clocortolone, Doxycycline, Pirlindole mesylate, Doxazosin, Deptropine,Nocodazole, Scopolamine, Oxybenzone, Halcinonide, Oxybutynin,Miconazole, Clomipramine, Cyproheptadine, Doxepin, Dyclonine,Salbutamol, Flavoxate, Amoxapine, Fenofibrate, Pimethixene and mixturesthereof.

The term “co-administration” or “combination therapy” is used todescribe a therapy in which at least two compounds in effective amountsare used to treat an autophagy mediated disease state or condition asotherwise described herein, either at the same time or within dosing oradministration schedules ascertainable by those of ordinary skill in theart. Although the term co-administration preferably includes theadministration of two active compounds to the patient at the same time,it is not necessary that the compounds be administered to the patient atthe same time, although effective amounts of the individual compoundswill be present in the patient at the same time. In addition, in certainembodiments, co-administration will refer to the fact that two compoundsare administered at significantly different times, but the effects ofthe two compounds are present at the same time. Thus, the termco-administration includes an administration in which one active agent(especially an autophagy modulator) is administered at approximately thesame time (contemporaneously), or from about one to several minutes toabout 24 hours or more than the other bioactive agent coadministeredwith the autophagy modulator. The additional bioactive agent may be anybioactive agent, but is generally selected from an additional autophagymediated compound, an additional anticancer agent, or another agent,such as a mTOR inhibitor such as pp242, rapamycin, envirolimus,everolimus or cidaforollimus, among others including epigallocatechingallate (EGCG), caffeine, curcumin or reseveratrol (which mTORinhibitors find particular use as enhancers of autophagy using thecompounds disclosed herein and in addition, in the treatment of cancerwith an autophagy modulator (inhibitor) as described herein, includingin combination with tetrachlorisophthalonitrile, phenylmercuric acetateand their pharmaceutically acceptable salts, which are inhibitors ofautophagy. It is noted that in the case of the treatment of cancer, theuse of an autophagy inhibitor is preferred, alone or in combination withan autophagy inducer (agonist) as otherwise described herein and/or amTOR inhibitor as described above. In certain embodiments, an mTORinhibitor selected from the group consisting of pp242, rapamycin,envirolimus, everolimus, cidaforollimus, epigallocatechin gallate(EGCG), caffeine, curcumin, reseveratrol and mixtures thereof may becombined with at least one agent selected from the group consisting ofdigoxin, xylazine, hexetidine and sertindole, the combination of suchagents being effective as autophagy modulators.

The term “cancer” is used throughout the specification to refer to thepathological process that results in the formation and growth of acancerous or malignant neoplasm, i.e., abnormal tissue that grows bycellular proliferation, often more rapidly than normal and continues togrow after the stimuli that initiated the new growth cease. Malignantneoplasms show partial or complete lack of structural organization andfunctional coordination with the normal tissue and most invadesurrounding tissues, metastasize to several sites, and are likely torecur after attempted removal and to cause the death of the patientunless adequately treated.

As used herein, the term neoplasia is used to describe all cancerousdisease states and embraces or encompasses the pathological processassociated with malignant hematogenous, ascitic and solid tumors.Representative cancers include, for example, stomach, colon, rectal,liver, pancreatic, lung, breast, cervix uteri, corpus uteri, ovary,prostate, testis, bladder, renal, brain/CNS, head and neck, throat,Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, leukemia,melanoma, non-melanoma skin cancer (especially basal cell carcinoma orsquamous cell carcinoma), acute lymphocytic leukemia, acute myelogenousleukemia, Ewing's sarcoma, small cell lung cancer, choriocarcinoma,rhabdomyosarcoma, Wilms' tumor, neuroblastoma, hairy cell leukemia,mouth/pharynx, esophagus, larynx, kidney cancer and lymphoma, amongothers, which may be treated by one or more compounds according to thepresent invention. In certain aspects, the cancer which is treated islung cancer, breast cancer, ovarian cancer and/or prostate cancer.

The term “tumor” is used to describe a malignant or benign growth ortumefacent.

The term “additional anti-cancer compound,” “additional anti-cancerdrug” or “additional anti-cancer agent” is used to describe any compound(including its derivatives) which may be used to treat cancer. The“additional anti-cancer compound,” “additional anti-cancer drug” or“additional anti-cancer agent” can be an anticancer agent which isdistinguishable from a CIAE-inducing anticancer ingredient such as ataxane, vinca alkaloid and/or radiation sensitizing agent otherwise usedas chemotherapy/cancer therapy agents herein. In many instances, theco-administration of another anti-cancer compound according to thepresent invention results in a synergistic anti-cancer effect. Exemplaryanti-cancer compounds for co-administration with formulations accordingto the present invention include anti-metabolites agents which arebroadly characterized as antimetabolites, inhibitors of topoisomerase Iand II, alkylating agents and microtubule inhibitors (e.g., taxol), aswell as tyrosine kinase inhibitors (e.g., surafenib), EGF kinaseinhibitors (e.g., tarceva or erlotinib) and tyrosine kinase inhibitorsor ABL kinase inhibitors (e.g. imatinib).

Anti-cancer compounds for co-administration include, for example,Aldesleukin; Alemtuzumab; alitretinoin; allopurinol; altretamine;amifostine; anastrozole; arsenic trioxide; Asparaginase; BCG Live;bexarotene capsules; bexarotene gel; bleomycin; busulfan intravenous;busulfan oral; calusterone; capecitabine; carboplatin; carmustine;carmustine with Polifeprosan 20 Implant; celecoxib; chlorambucil;cisplatin; cladribine; cyclophosphamide; cytarabine; cytarabineliposomal; dacarbazine; dactinomycin; actinomycin D; Darbepoetin alfa;daunorubicin liposomal; daunorubicin, daunomycin; Denileukin diftitox,dexrazoxane; docetaxel; doxorubicin; doxorubicin liposomal;Dromostanolone propionate; Elliott's B Solution; epirubicin; Epoetinalfa estramustine; etoposide phosphate; etoposide (VP-16); exemestane;Filgrastim; floxuridine (intraarterial); fludarabine; fluorouracil(5-FU); fulvestrant; gemtuzumab ozogamicin; gleevec (imatinib);goserelin acetate; hydroxyurea; Ibritumomab Tiuxetan; idarubicin;ifosfamide; imatinib mesylate; Interferon alfa-2a; Interferon alfa-2b;irinotecan; letrozole; leucovorin; levamisole; lomustine (CCNU);meclorethamine (nitrogen mustard); megestrol acetate; melphalan (L-PAM);mercaptopurine (6-MP); mesna; methotrexate; methoxsalen; mitomycin C;mitotane; mitoxantrone; nandrolone phenpropionate; Nofetumomab; LOddC;Oprelvekin; oxaliplatin; paclitaxel; pamidronate; pegademase;Pegaspargase; Pegfilgrastim; pentostatin; pipobroman; plicamycin;mithramycin; porfimer sodium; procarbazine; quinacrine; Rasburicase;Rituximab; Sargramostim; streptozocin; surafenib; talbuvidine (LDT);talc; tamoxifen; tarceva (erlotinib); temozolomide; teniposide (VM-26);testolactone; thioguanine (6-TG); thiotepa; topotecan; toremifene;Tositumomab; Trastuzumab; tretinoin (ATRA); Uracil Mustard; valrubicin;valtorcitabine (monoval LDC); vinblastine; vinorelbine; zoledronate; andmixtures thereof, among others.

Co-administration of one of the formulations of the invention withanother anticancer agent will often result in a synergistic enhancementof the anticancer activity of the other anticancer agent, an unexpectedresult. One or more of the present formulations comprising an autophagymodulator may also be co-administered with another bioactive agent(e.g., antiviral agent, antihyperproliferative disease agent, agentswhich treat chronic inflammatory disease, among others as otherwisedescribed herein).

The term “antiviral agent” refers to an agent which may be used incombination with autophagy modulators as otherwise described herein totreat viral infections, including HIV infections, HBV infections and/orHCV infections. Exemplary anti-HIV agents include, for example,nucleoside reverse transcriptase inhibitors (NRTI), non-nucleosidereverse transcriptase inhibitors (NNRTI), protease inhibitors, fusioninhibitors, among others, exemplary compounds of which may include, forexample, 3TC (Lamivudine), AZT (Zidovudine), (-)-FTC, ddI (Didanosine),ddC (zalcitabine), abacavir (ABC), tenofovir (PMPA), D-D4FC (Reverset),D4T (Stavudine), Racivir, L-FddC, L-FD4C, NVP (Nevirapine), DLV(Delavirdine), EFV (Efavirenz), SQVM (Saquinavir mesylate), RTV(Ritonavir), IDV (Indinavir), SQV (Saquinavir), NFV (Nelfinavir), APV(Amprenavir), LPV (Lopinavir), fusion inhibitors such as T20, amongothers, fuseon and mixtures thereof, including anti-HIV compoundspresently in clinical trials or in development. Exemplary anti-HBVagents include, for example, hepsera (adefovir dipivoxil), lamivudine,entecavir, telbivudine, tenofovir, emtricitabine, clevudine,valtoricitabine, amdoxovir, pradefovir, racivir, BAM 205, nitazoxanide,UT 231-B, Bay 41-4109, EHT899, zadaxin (thymosin alpha-1) and mixturesthereof. Anti-HCV agents include, for example, interferon, pegylatedintergeron, ribavirin, NM 283, VX-950 (telaprevir), SCH 50304, TMC435,VX-500, BX-813, SCH503034, R1626, ITMN-191 (R7227), R7128, PF-868554,TT033, CGH-759, GI 5005, MK-7009, SIRNA-034, MK-0608, A-837093, GS 9190,ACH-1095, GSK625433, TG4040 (MVA-HCV), A-831, F351, NSSA, NS4B, ANA598,A-689, GNI-104, IDX102, ADX184, GL59728, GL60667, PSI-7851, TLR9Agonist, PHX1766, SP-30 and mixtures thereof.

The term “compound” is used herein to refer to any specific chemicalcompound disclosed herein.

In certain non-limiting embodiments, an increase or a decrease in asubject or test sample of the level of measured protein or geneexpression or autophagic change as compared to a comparable level ofmeasured protein or gene expression or autophagic change in a controlsubject or sample can be an increase or decrease in the magnitude ofapproximately ±5,000-10,000%, or approximately ±2,500-5,000%, orapproximately ±1,000-2,500%, or approximately ±500-1,000%, orapproximately ±250-500%, or approximately ±100-250%, or approximately±50-100%, or approximately ±25-50%, or approximately ±10-25%, orapproximately ±10-20%, or approximately ±10-15%, or approximately±5-10%, or approximately ±1-5%, or approximately ±0.5-1%, orapproximately ±0.1-0.5%, or approximately ±0.01-0.1%, or approximately±0.001-0.01%, or approximately ±0.0001-0.001%.

The values obtained from controls are reference values representing aknown health status and the values obtained from test samples orsubjects are reference values representing a known disease status. Theterm “control,” as used herein, can mean a sample of preferably the samesource (e.g. blood, serum, tissue etc.) which is obtained from at leastone healthy subject to be compared to the sample to be analyzed. Inorder to receive comparable results the control as well as the sampleshould be obtained, handled and treated in the same way. In certainexamples, the number of healthy individuals used to obtain a controlvalue may be at least one, preferably at least two, more preferably atleast five, most preferably at least ten, in particular at least twenty.However, the values may also be obtained from at least one hundred, onethousand or ten thousand individuals.

The term “patient” or “subject” refers to an animal, such as a mammal,or a human, in need of treatment or therapy to which compounds accordingto the present invention are administered in order to treat a conditionor disease state associated with autophagy.

The term “sample” encompasses a variety of sample types obtained from anorganism and can be used in a diagnostic or monitoring assay. The termencompasses blood and/or plasma and other liquid samples of biologicalorigin, solid tissue samples, such as a biopsy specimen or tissuecultures or cells derived therefrom and the progeny thereof. The termencompasses samples that have been manipulated in any way after theirprocurement, such as by treatment with reagents, solubilization, orenrichment for certain components. The term encompasses a clinicalsample, and also includes cells in cell culture, cell supernatants, celllysates, serum, plasma, biological fluids, and tissue samples.

The term “body fluid” refers to a biological sample of liquid from amammal, e.g., from a human. Such fluids include aqueous fluids such asserum, plasma, lymph fluid, synovial fluid, follicular fluid, seminalfluid, amniotic fluid, milk, whole blood, urine, cerebrospinal fluid,saliva, sputum, tears, perspiration, mucus, tissue culture medium,tissue extracts, and cellular extracts. Particular bodily fluids thatare interest in the context of the present invention include serum,plasma, and blood.

According to various embodiments, the compounds according to the presentinvention may be used for treatment or prevention purposes in the formof a pharmaceutical composition. This pharmaceutical composition maycomprise one or more of compound, or active ingredient, as describedherein.

The pharmaceutical composition may also comprise a pharmaceuticallyacceptable excipient, additive or inert carrier. The pharmaceuticallyacceptable excipient, additive or inert carrier may be in a form chosenfrom a solid, semi-solid, and liquid. The pharmaceutically acceptableexcipient or additive may be chosen from a starch, crystallinecellulose, sodium starch glycolate, polyvinylpyrolidone,polyvinylpolypyrolidone, sodium acetate, magnesium stearate, sodiumlauryl sulfate, sucrose, gelatin, silicic acid, polyethylene glycol,water, alcohol, propylene glycol, vegetable oil, corn oil, peanut oil,olive oil, surfactants, lubricants, disintegrating agents, preservativeagents, flavoring agents, pigments, and other conventional additives.The pharmaceutical composition may be formulated by admixing the activewith a pharmaceutically acceptable excipient or additive.

The pharmaceutical composition may be in a form chosen from sterileisotonic aqueous solutions, pills, drops, pastes, cream, spray(including aerosols), capsules, tablets, sugar coating tablets,granules, suppositories, liquid, lotion, suspension, emulsion, ointment,gel, and the like. Administration route may be chosen from subcutaneous,intravenous, intestinal, parenteral, oral, buccal, nasal, intramuscular,transcutaneous, transdermal, intranasal, intraperitoneal, and topical.The pharmaceutical compositions may be immediate release,sustained/controlled release, or a combination of immediate release andsustained/controlled release depending upon the compound(s) to bedelivered, the compound(s), if any, to be coadministered, as well as thedisease state and/or condition to be treated with the pharmaceuticalcomposition. A pharmaceutical composition may be formulated withdiffering compartments or layers in order to facilitate effectiveadministration of any variety consistent with good pharmaceuticalpractice.

The subject or patient may be chosen from, for example, a human, amammal such as domesticated animal, or other animal. The subject mayhave one or more of the disease states, conditions or symptomsassociated with autophagy as otherwise described herein.

The compounds according to the present invention may be administered inan effective amount to treat or reduce the likelihood of anautophagy-mediated disease and/or condition as well one or more symptomsassociated with the disease state or condition. One of ordinary skill inthe art would be readily able to determine an effective amount of activeingredient by taking into consideration several variables including, butnot limited to, the animal subject, age, sex, weight, site of thedisease state or condition in the patient, previous medical history,other medications, etc.

For example, the dose of an active ingredient which is useful in thetreatment of an autophagy mediated disease state, condition and/orsymptom for a human patient is that which is an effective amount and mayrange from as little as 100 μg or even less to at least about 500 mg ormore, which may be administered in a manner consistent with the deliveryof the drug and the disease state or condition to be treated. In thecase of oral administration, active is generally administered from oneto four times or more daily. Transdermal patches or other topicaladministration may administer drugs continuously, one or more times aday or less frequently than daily, depending upon the absorptivity ofthe active and delivery to the patient's skin. Of course, in certaininstances where parenteral administration represents a favorabletreatment option, intramuscular administration or slow IV drip may beused to administer the pharmaceutical composition. The amount of activeingredient which is administered to a human patient preferably rangesfrom about 0.05 mg/kg to about 10 mg/kg, about 0.1 mg/kg to about 7.5mg/kg, about 0.25 mg/kg to about 6 mg/kg, about 1.25 to about 5.7 mg/kg.

The compound according to the present invention may be administered atthe first signs of the onset of an autophagy mediated disease state,condition or symptom. For example, the compound may be administered forthe purpose of lung or heart function and/or treating or reducing thelikelihood of any one or more of the disease states or conditions whichbecome manifest during an inflammation-associated metabolic disorder ortuberculosis or associated disease states or conditions, including pain,high blood pressure, renal failure, or lung failure. The dose of activeingredient may be administered at the first sign of relevant symptomsprior to diagnosis, in anticipation of the disease or disorder, or inanticipation of decreased bodily function or any one or more of theother symptoms or secondary disease states or conditions associated withan autophagy mediated disorder to condition.

These and other aspects of the invention are described further in thefollowing illustrative examples.

EXAMPLES Example 1

Preparation of3-(4-methoxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione (BPY-103),3 ml p-methoxybenzaldehyde (24.7 mmoles), 6.8 milliliters (ml)2-methoxyacetophenoe (49.4 mmoles), 20 ml of reagent alcohol (90%ethanol, 5% methanol, 5% isopropanol), and a stir bar was added to a 50ml round-bottom flask. 3.3 g of KOH (85%) in 15 ml of water was thenadded to the flask, which caused the reaction mixture to become opaquewithin 5 minutes. The flask was then fitted with a septum and heatedunder nitrogen in a 105° C. sand bath for 12 hours. After this time, thecrude product was precipitated in 200 ml of water before filtering andwashing with methanol. The pure product was then recrystallized frommethanol, yielding 2.34 grams (g) of3-(4-methoxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione. A furtherrecrystallization from the mother liquor yielded a further 1.10 g of3-(4-methoxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione. The totalyield was 3.44 g (33.3%)

¹H NMR (90 MHz, CDCl3): δ=7.55-6.85 (m, 12H, aromatics), 3.932 (s, 1H),3.882 (s, 6H, peripheral methoxy groups), 3.788 (s, 3H, central methoxygroups), 3.412 (2H, CH₂), 3.332 (2H, CH₂). Melting range: 95-97° C.

Example 2 Preparation of3-(4-allyloxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione(BPY-101A)

0.93 g (4.89 mmoles) of BPY-101B (described in Example 2), 10 ml reagentalcohol (90% ethanol, 5% methanol, 5% isopropanol), a stir bar, and 1.6ml 2-methoxyacetophenone was added to a 50 ml round-bottom flask. Theflask was purged with N₂ for 5 min, and 0.67 g of 85% KOH in 10 ml ofwater was added, which caused immediate precipitation. More reagentalcohol was added until the precipitates were re-dissolved, giving asolution with a strong yellow color. The flask was then placed in a 105°C. sand bath for 19 hours, while the contents were stirred under anitrogen atmosphere. After this time, the reaction mixture wasprecipitated into 300 ml of pH≅4 water, and the solids filtered andrecrystallized from methanol. 1.08 g of the compound was recovered(49.7%)

¹H NMR (90 MHz, CDCl₃): δ=7.618-6.811 (m, 12H, aromatics), 6.042 (m, 1H,allyl CH₂-CH=CH₂), 5.564 (dd, 1H, allyl CH₂-CH=CH ₂) 5.339 (dd, 1H,allyl CH₂-CH=CH ₂) 3.972 (1 h, partially obscured), 3.926 (s, 6H,methoxy), 3.461 (2H, CH ₂), 3.382 (2H, CH ₂). Melting range: 86-88° C.

Example 3 Preparation of3-(4-hydroxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione (BPY-101)

1.09 g BPY-101A (described in Example 3), 15 ml methanol, and 0.46 gsodium tert-butoxide was added to a 50 ml round bottomed flask. Themixture was sparged with N₂ and stirred for 15 minutes. After this time,0.46 g tetrakis(triphenylphosphine)palladium(0) was added to thereaction mixture and stirred under N₂ with occasional heating tore-dissolve particulates. After 1 hour, only one spot on the baselineremained in TLC (CHCl₃). The reaction mixture was added to 100 ml pH≅12water and washed with 2×100 CHCl₃. The water phase was then acidified(pH≅4) with HCl, and extracted with 3×300 ml CHCl₃. The organic phasewas then dried over Na₂SO₄ and reduced in vaccuo leaving an orangesolid.

¹H NMR (90 MHz, CDCl₃): δ=7.574-6.645 (m, 12H, aromatics), 3.897(s, 6H,methoxy), 3.429 (2H, CH ₂), 3.351 (2H, CH ₂). Note: The single protonpeak could not be definitely assigned and is most likely overlapped bythe methoxy peaks.

Example 4 Preparation of 2,6-di(2-methoxyphenyl)-4-(4-methoxyphenyl)pyridine (BPY-104)

Into a 50 ml round-bottomed flask was added 0.52 g of3-(4-methoxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione (BPY-103),20 ml of glacial acetic acid, a stir bar and 3.1 g ammonium acetate. Theflask was fitted with a reflux condenser and the solution was refluxedwhile stirring for 4 h. The flask was then removed from heat and thecrude product was precipated into 300 ml of water. The pure product wasobtained by recrystallization from a mixture of chloroform and methanol.Formation of the product was confirmed by the disappearance of the CH₂peaks at 3.412 ppm and 3.332 ppm in the NMR spectrum, and also by thedramatic increase in melting range. ¹H NMR (90 MHz, CDCl₃): δ=7.95-6.75m, 14H, aromatic), 3.9-3.7(9H, methoxy). Melting range: 188-190° C.

Although the present invention has been described in considerable detailwith reference to certain preferred embodiments, other embodiments arepossible. The steps disclosed for the present methods, for example, arenot intended to be limiting nor are they intended to indicate that eachstep is necessarily essential to the method, but instead are exemplarysteps only. Therefore, the scope of the appended claims should not belimited to the description of preferred embodiments contained in thisdisclosure. All references cited herein are incorporated by reference intheir entirety.

FEATURES OF THE INVENTION

1. A compound having the following structural formula:

2. A compound having the following structural formula:

3. A compound having the following structural formula:

4. A compound having the following structural formula:

5. A compound having the following structural formula:

6. A compound having the following structural formula:

7. A compound having the following structural formula:

8. A compound having the following structural formula:

9. A compound having the following structural formula:

10. A compound having the following structural formula:

11. A compound having the following structural formula:

12. A compound having the following structural formula:

13. A compound having the following structural formula:

14. A compound having the following structural formula:

15. A compound having the following structural formula:

16. A compound having the following structural formula:

17. A compound having the following structural formula:

18. A compound having the following structural formula:

19. A compound having the following structural formula:

20. A compound having the following structural formula:

21. A method of using the compound of any of features 1-20 to treat anautophagy-mediated disease state or condition.

22. The method of feature 21, wherein the autophagy-mediated diseasestate or condition is cancer, lysosomal storage diseases, Alzheimer'sdisease, Parkinson's disease and other ataxias such as Huntington'sdisease; a chronic inflammatory disease such as Crohn's disease,diabetes I, diabetes II, metabolic syndrome, an inflammation-associatedmetabolic disorder, liver disease, renal disease, cardiovasculardisease, muscle degeneration and atrophy, frailty in aging, spinal cordinjury, infectious disease, chronic pain, depression related syndromes,and developmental disease.

23. The method according to feature 22 wherein the cancer is stomach,colon, rectal, liver, pancreatic, lung, breast, cervix uteri, corpusuteri, ovary, prostate, testis, bladder, renal, brain/CNS, head andneck, throat, Hodgkin's disease, non-Hodgkin's lymphoma, multiplemyeloma, leukemia, melanoma, non-melanoma skin cancer, acute lymphocyticleukemia, acute myelogenous leukemia, Ewing's sarcoma, small cell lungcancer, choriocarcinoma, rhabdomyosarcoma, Wilms' tumor, neuroblastoma,hairy cell leukemia, mouth/pharynx, esophagus, larynx, kidney cancer orlymphoma.

24. The method according to feature 22 wherein the chronic inflammatorydisease is inflammatory bowel disease, rheumatoid arthritis, lupus,multiple sclerosis, chronic obstructive pulmonary disease/COPD,pulmonary fibrosis, cystic fibrosis or Sjogren's disease.

25. The method according to feature 22 wherein the cardiovasculardisease is ischemia, stroke, pressure overload, complications duringreperfusion and arteriosclerosis.

26. The method according to feature 22 wherein the infectious disease isa viral infection or a secondary condition of the viral infection.

27. The method according to feature 26 wherein the viral infection isHIV (I and or II) and said secondary condition is AIDS, hepatitis Bvirus (HBV) or hepatitis C virus, influenza virus and herpes virus.

28. The method according to feature 22 wherein said lysosomal storagedisease is activator deficiency/GM2 gangliosidosis, alpha-mannosidosis,aspartylglucoaminuria, cholesteryl ester storage disease, chronichexosaminidase A deficiency, cystinosis, Danon disease, Fabry disease,Farber disease, fucosidosis, galactosialidosis, Gaucher Disease (TypesI, II and III), GM1 Ganliosidosis, including infantile, lateinfantile/juvenile and adult/chronic), Hunter syndrome (MPS II), I-Celldisease/Mucolipidosis II, Infantile Free Sialic Acid Storage Disease(ISSD), Juvenile Hexosaminidase A Deficiency, Krabbe disease, Lysosomalacid lipase deficiency, Metachromatic Leukodystrophy, Hurler syndrome,Scheie syndrome, Hurler-Scheie syndrome, Sanfilippo syndrome, MorquioType A and B, Maroteaux-Lamy, Sly syndrome, mucolipidosis, multiplesulfate deficiency, Niemann-Pick disease, Neuronal ceroidlipofuscinoses, CLN6 disease, Jansky-Bielschowsky disease, Pompedisease, pycnodysostosis, Sandhoff disease, Schindler disease, Tay-Sachsor Wolman disease.

29. A pharmaceutical composition comprising:

-   -   (a) the compound of features 1-20 in an effective amount; and        optionally    -   (b) a pharmaceutically-acceptable carrier, additive and/or        excipient, and further optionally;    -   (c) at least one additional bioactive agent.    -   or a secondary condition of the viral infection.

30. The composition according to feature 29 wherein the additionalautophagy modulator is selected from the group consisting ofbenzethonium, niclosamide, monensin, bromperidol, levobunolol,dehydroisoandosterone 3-acetate, sertraline, tamoxifen, reserpine,hexachlorophene, dipyridamole, harmaline, prazosin, lidoflazine,thiethylperazine, dextromethorphan, desipramine, mebendazole, canrenone,chlorprothixene, maprotiline, homochlorcyclizine, loperamide,nicardipine, dexfenfluramine, nilvadipine, dosulepin, biperiden,denatonium, etomidate, toremifene, tomoxetine, clorgyline, zotepine,beta-escin, tridihexethyl, ceftazidime, methoxy-6-harmalan,melengestrol, albendazole, rimantadine, chlorpromazine, pergolide,cloperastine, prednicarbate, haloperidol, clotrimazole, nitrofural,iopanoic acid, naftopidil, methimazole, trimeprazine, ethoxyquin,clocortolone, doxycycline, pirlindole mesylate, doxazosin, deptropine,nocodazole, scopolamine, oxybenzone, halcinonide, oxybutynin,miconazole, clomipramine, cyproheptadine, doxepin, dyclonine,salbutamol, flavoxate, amoxapine, fenofibrate, pimethixene,pharmaceutically acceptable salts thereof and mixtures thereof.

16. The composition according to feature 29 wherein the additionalbioactive agent is an antibiotic or an antiviral agent.

17. The composition according to feature 29 wherein the bioactive agentincludes an anticancer agent.

What is claimed is:
 1. A compound having the following backbonestructural formula:


2. The compound of claim 1, wherein the compound is selected from thegroup consisting of:N-(3,4-dimethoxypheny)-1,8-Dioxa-3-aza-2,4-dihydro-2H-5-phenyl-anthracen-7-one,N-(3,4-dimethoxybenzyl)-1,8-Dioxa-3-aza-2,4-dihydro-2H-5-methyl-anthracen-7-one,N-benzyl-1,8-Dioxa-3-aza-3,4dihydro-2H-anthracen-7-one series, andN-(3,4-dimethoxypheny)-1,8-Dioxa-3-aza-2,4-dihydro-2H-5-methyl-anthracen-7-one.3. A compound having the following backbone structural formula:


4. The compound of claim 3, wherein the compound is selected from thegroup consisting of:3-(4-methoxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione,3-(4-hydroxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione,3-(4-methoxyphenyl)-1,5-bis (2-methoxyphenyl)-1,5-pentanedione,3-(4-allyloxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione, 3-(4-hydroxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione,3-(3,4,5-trimethoxymethoxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione,3-(4-allyloxyphenyl)-1,5-bis(2-methoxyphenyl)-1,5-pentanedione, and3-(4-dimethylaminophenyl)-1,5- bis(2-methoxyphenyl)-1,5-pentanedione. 5.A method of using the compound of claim 2 to treat an autophagy-mediateddisease state or condition.
 6. A method of using the compound of claim 4to treat an autophagy-mediated disease state or condition.
 7. The methodclaim 5, wherein the autophagy-mediated disease state or condition iscancer, lysosomal storage diseases, Alzheimer's disease, Parkinson'sdisease and other ataxias such as Huntington's disease; a chronicinflammatory disease such as Crohn's disease, diabetes I, diabetes II,metabolic syndrome, an inflammation-associated metabolic disorder, liverdisease, renal disease, cardiovascular disease, muscle degeneration andatrophy, frailty in aging, spinal cord injury, infectious disease,chronic pain, depression related syndromes, and developmental disease.8. The method claim 6, wherein the autophagy-mediated disease state orcondition is cancer, lysosomal storage diseases, Alzheimer's disease,Parkinson's disease and other ataxias such as Huntington's disease; achronic inflammatory disease such as Crohn's disease, diabetes I,diabetes II, metabolic syndrome, an inflammation-associated metabolicdisorder, liver disease, renal disease, cardiovascular disease, muscledegeneration and atrophy, frailty in aging, spinal cord injury,infectious disease, chronic pain, depression related syndromes, anddevelopmental disease.
 9. The composition of claim 2 wherein thecomposition further comprises an autophagy modulator selected from thegroup comprising benzethonium, niclosamide, monensin, bromperidol,levobunolol, dehydroisoandosterone 3-acetate, sertraline, tamoxifen,reserpine, hexachlorophene, dipyridamole, harmaline, prazosin,lidoflazine, thiethylperazine, dextromethorphan, desipramine,mebendazole, canrenone, chlorprothixene, maprotiline,homochlorcyclizine, loperamide, nicardipine, dexfenfluramine,nilvadipine, dosulepin, biperiden, denatonium, etomidate, toremifene,tomoxetine, clorgyline, zotepine, beta-escin, tridihexethyl,ceftazidime, methoxy-6-harmalan, melengestrol, albendazole, rimantadine,chlorpromazine, pergolide, cloperastine, prednicarbate, haloperidol,clotrimazole, nitrofural, iopanoic acid, naftopidil, methimazole,trimeprazine, ethoxyquin, clocortolone, doxycycline, pirlindolemesylate, doxazosin, deptropine, nocodazole, scopolamine, oxybenzone,halcinonide, oxybutynin, miconazole, clomipramine, cyproheptadine,doxepin, dyclonine, salbutamol, flavoxate, amoxapine, fenofibrate,pimethixene, pharmaceutically acceptable salts thereof and mixturesthereof.
 10. The composition of claim 9, wherein the additionalbioactive agent is an antibiotic, an antiviral agent, or an anticanceragent.
 11. A pharmaceutical composition comprising the compound of claim2 in an effective amount.
 12. The composition of claim 11, furthercomprising a pharmaceutically-acceptable carrier, additive or excipient.13. The composition of claim 12, further comprising at least oneadditional bioactive agent or a secondary condition of the viralinfection.
 14. The composition of claim 4 wherein the compositionfurther comprises an autophagy modulator selected from the groupcomprising benzethonium, niclosamide, monensin, bromperidol,levobunolol, dehydroisoandosterone 3-acetate, sertraline, tamoxifen,reserpine, hexachlorophene, dipyridamole, harmaline, prazosin,lidoflazine, thiethylperazine, dextromethorphan, desipramine,mebendazole, canrenone, chlorprothixene, maprotiline,homochlorcyclizine, loperamide, nicardipine, dexfenfluramine,nilvadipine, dosulepin, biperiden, denatonium, etomidate, toremifene,tomoxetine, clorgyline, zotepine, beta-escin, tridihexethyl,ceftazidime, methoxy-6-harmalan, melengestrol, albendazole, rimantadine,chlorpromazine, pergolide, cloperastine, prednicarbate, haloperidol,clotrimazole, nitrofural, iopanoic acid, naftopidil, methimazole,trimeprazine, ethoxyquin, clocortolone, doxycycline, pirlindolemesylate, doxazosin, deptropine, nocodazole, scopolamine, oxybenzone,halcinonide, oxybutynin, miconazole, clomipramine, cyproheptadine,doxepin, dyclonine, salbutamol, flavoxate, amoxapine, fenofibrate,pimethixene, pharmaceutically acceptable salts thereof and mixturesthereof.
 15. The composition of claim 14, wherein the additionalbioactive agent is an antibiotic, an antiviral agent, or an anticanceragent.
 16. A pharmaceutical composition comprising the compound of claim4 in an effective amount.
 17. The composition of claim 16, furthercomprising a pharmaceutically-acceptable carrier, additive or excipient.18. The composition of claim 17, further comprising at least oneadditional bioactive agent or a secondary condition of the viralinfection.